The heat shock response (HSR) induced by cell stress is a common protection mechanism elicited in a wide range of tumors. This response is closely associated with a poor prognosis and resistance to therapy. Phase I of this proposed research aims to identify drug targets involved in the HSR. These potential drugs, which are expected to inhibit the HSR, may be used in combination with known activators of cell stress, such as heat shock, Bortezomib and Geldanamycin, for improved treatment of cancers. In addition, completion of Phase I of the proposed research will result in generation of a prototype database of HSR pathway regulation in a wide range of cell lines of diverse tumor origin. Phase II of this research proposes in-depth characterization of drug targets involved in the HSR/PTS pathway and development of the reporter cell lines for high throughput screening (HTS) of a chemical compound library. Given our unique interface with cellbased drug screening facilities available at Roswell Park Cancer Center, we expect to develop a pipeline of potential drugs with fast conversion of pathway knowledge into efficient strategies of drug identification. At completion of the proposed studies, we anticipate that we will have generated a novel class of anti-tumor drugs with no equivalent properties previously described in the literature or applied in clinical trials. We propose that these drugs be used in combination with drugs known to activate the HSR through inhibition of basal heat shock machinery, such as Bortezomib and Geldanamicin, or in combination with heat therapy. Our preliminary studies argue in favor of a very strong synergistic effect of these drugs in the induction of tumor cell death. PUBLIC HEALTH RELEVANCE: The goal of this project is to identify drug targets within the heat shock response pathway for establishment of novel approaches to discover anti-cancer therapies through isolation of chemical compounds blocking heat shock response in tumors. Drug targets will be identified through RNAi technology, allowing for development of RNAi library products and generation of databases of cell lines, genes, and molecular pathways.